关节软骨与软骨下骨改变在不同骨关节炎动物模型中的特点

目的 探讨不同骨关节炎(osteoarthritis,OA)动物模型中软骨下骨和关节软骨的病理改变特征。方法 采用三种SD大鼠骨关节炎模型,将24只6月龄雌性大鼠随机分为4组:假手术对照组(Sham,n=6),前交叉韧带切除手术组(ACLT,n=6),木瓜蛋白酶关节腔注射组(Papain,n=6),以及卵巢切除术组(OVX,n=6)。造模后8周取膝关节,胫骨平台行Micro-CT扫描分析,关节软骨行甲苯胺蓝染色、Mankin评分,比较软骨下骨和关节软骨改变情况。结果 造模操作后8周,不同OA模型的软骨破坏程度有所不同。ACLT和Papain组软骨破坏比较严重,OVX组软骨变化较轻。所有OA模型中的软骨下松质骨均发生改变,OVX组相对于Sham对照组,软骨下骨微结构显著疏松,而ACLT组与Papain组相对于Sham对照组,软骨下骨微结构没有显著改变,但相对于OVX组,有显著性差异。三种OA模型的软骨下骨板厚度都较Sham组减少。结论 三种动物模型软骨下骨和关节软骨都发生明显病理改变,并且改变有所不同。不同OA模型代表不同病理,预示着软骨下骨所发挥的作用有所不同,这为进一步研究不同类型OA发生发展的机制,以及将软骨下骨作为OA治疗的可能靶点提供了更多的依据。     

葛根异黄酮对去卵巢大鼠骨骼生物力学的影响

目的:观察葛根异黄酮对去卵巢大鼠骨骼生物力学的影响。方法:选用健康3月龄SD雌性大鼠50只,随机分为5组:假手术组、模型对照组、低、中、高剂量葛根异黄酮组;假手术组切除卵巢旁脂肪组织,其余各组均切除双侧卵巢,建立去卵巢大鼠动物模型。按25、50、100 mg/kg 3个剂量葛根异黄酮分别给去卵巢大鼠灌胃6个月,并于实验结束后处死大鼠,检测腰椎骨骼生物力学功能、骨矿含量和股骨骨钙含量及子宫重量。结果:高剂量葛根异黄酮治疗组6个月后腰椎最大负荷、结构强度和骨矿含量均较模型对照组明显改善,股骨骨钙含量也明显增高;各个葛根异黄酮治疗组子宫重量均较模型对照组明显增加。结论:葛根异黄酮可改善去卵巢大鼠骨骼生物力学功能,具有雌激素样的抗骨质疏松作用。     

胰岛素信号通路相关因子在2型糖尿病合并骨质疏松大鼠肝组织的表达

通过高糖高脂饲料联合小剂量链脲佐菌素和去卵巢手术制备2型糖尿病合并骨质疏松大鼠模型,探讨2型糖尿病合并骨质疏松大鼠肝组织胰岛素信号通路相关因子的表达及意义。结果表明:随着时间延长,2型糖尿病合并骨质疏松组(DOVX组)肝组织IGF-1、IRS-1较其他组mRNA及蛋白表达减少,单纯去卵巢组(NOVX组)IGF-1、IRS-1 mRNA及蛋白表达较假手术对照组(NS组)降低;糖尿病组(DS组)IRS-2较NS组mRNA及蛋白表达下降,但NOVX组与NS组IRS-2 mRNA及蛋白表达比较无明显差别。以上结果表明,2型糖尿病合并骨质疏松的发生可能与肝脏胰岛素信号通路受抑制有关。     

ERα基因过表达对去卵巢骨质疏松小鼠骨密度及钙磷代谢的影响

目的:探讨雌激素受体α(ERα)基因过表达对去卵巢骨质疏松小鼠骨代谢及钙磷代谢的影响,为靶向基因治疗骨质疏松症提供实验依据.方法:选择SPF级雌性小鼠30只,随机分为假手术组、模型组和ERα过表达组,每组10只.采用双侧卵巢切除法制备绝经后骨质疏松小鼠模型,模型建立完成后,ERα过表达组通过椎体髓腔注射的方法转染携带小...     

仙灵骨葆对骨质疏松大鼠OPG/RANK/RANKL 表达的影响

目的:观察仙灵骨葆治疗骨质疏松模型大鼠后,对大鼠体内OPG/RANKL/RANK表达的影响。方法:卵巢摘除法建立SD大鼠骨质疏松模型,设立假手术组、对照组(单纯去卵巢组)、雌激素组(给予17β-雌二醇)和治疗组(给予仙灵骨葆)。术后1周开始给药,给药12周后检测各组大鼠股骨骨密度,ELISA法检测血清中OPG/RANKL含量,RT-PCR检测骨组织中OPG/RANK/RAN-KL mRNA表达,免疫组化检测骨组织中RANK的表达。结果:对照组大鼠骨密度显著低于假手术组;治疗组和雌激素组大鼠O-PG表达显著高于对照组,RANK及RANKL的表达显著低于对照组。结论:采用卵巢摘除法成功建立大鼠骨质疏松模型;仙灵骨葆可促进骨质疏松大鼠OPG的表达,并抑制RANK及RANKL的表达,对骨质疏松模型大鼠有治疗作用。     

金雀异黄酮对骨质疏松大鼠体内Sost表达的影响

目的:观察金雀异黄酮对去卵巢大鼠骨质疏松发生的影响并探讨其分子机制.方法:将60只雌性SD大鼠随机分为:空白对照组、模型对照组、尼尔雌醇组、金雀异黄酮高(18 mg/kg)、中(9 mg/kg)、低(4.5 mg/kg)剂量组.除空白对照组外,其余大鼠切除双侧卵巢诱发骨质疏松.模型复制成功后,给予含有不同剂量金雀异黄酮的饲料喂食骨质疏松大鼠.12周后行股骨干骨密度测定并行RT-PCR和Western blot检测股骨组织中sost基因表达水平的变化.结果:和模型对照组相比,中剂量金雀异黄酮组大鼠骨密度显著增加(P<0.01)股骨组织中sost表达明显下调.结论:金雀异黄酮对骨质疏松大鼠有治疗作用,该作用可能与下调sost表达有关.     

补肾化痰方对去卵巢大鼠血清TRACP5b 和β-CTX 的影响

目的:观察补肾化痰方对去卵巢骨质疏松症大鼠血清脂联素抗酒石酸酸性磷酸酶5b(tartarte-resistant acid phosphatase 5b,TRACP5b)和血清β-I型胶原C-末端肽(C-terminal telopeptides of type I collagen,β-CTX)水平的影响。方法:SD大鼠行双侧卵巢切除术;术后3月以中剂量和高剂量补肾化痰方灌胃给药;术后5月,双能X线骨密度测定仪检测大鼠股骨骨密度,ELISA法检测大鼠血清TRACP5b和β-CTX含量。结果:补肾化痰方中、高剂量组均能显著提高去卵巢骨质疏松症大鼠的骨密度,降低血清TRACP5b和β-CTX水平。结论:补肾化痰方对去卵巢大鼠骨质疏松症有较好的治疗作用。     

动态观察去卵巢骨质疏松大鼠股骨骨微结构的变化

目的:绝经后骨质疏松是好发于中老年女性人群中的骨代谢疾病,去卵巢骨质疏松大鼠模型是国内外通用的模拟绝经后骨质疏松发生的经典动物模型,本研究通过观察去卵巢骨质疏松大鼠股骨骨微结构的动态变化,为骨质疏松大鼠模型的临床应用提供理论参考依据。方法:将90只3月龄雌性SD大鼠按体重分层后随机分为基础组(10只)、假手术组(40只)和去卵巢组(40只)。分别在手术前(基础组)和后的3、6、12、24周,腹主动脉取血处死基础组以及假手术组和去卵巢组大鼠,每组各8-10只。每组中随机取6只大鼠,对其左股骨行micro-CT扫描及三维结构重建。选择股骨远端距生长板远端1 mm处,2.0 mm×3.5 mm,厚0.9 mm的骨组织为感兴趣区域,对感兴趣区域进行骨形态计量学分析。结果:与0周组比较,从去卵巢3周开始一直持续到24周,去卵巢组大鼠股骨vBMD、BV/TV和Tb.N显著降低,Tb.Sp和SMI显著升高,而Tb.Th无显著变化;与0周组比较,从假手术后3周开始一直到24周,假手术组所有检测指标均无显著变化。与同周龄假手术组比较,从去卵巢3周开始一直持续到24周,去卵巢组大鼠股骨Tb.N、BV/TV和vBMD显著降低,Tb.Sp显著升高,而Tb.Th没有显著变化。从去卵巢6周开始一直到24周,去卵巢组大鼠SMI显著增加。结论:3月龄大鼠股骨远端的骨微结构在去卵巢3周时就出现显著变化。提示,采用3月龄大鼠进行抗骨质疏松药物筛选时,去卵巢3周后就可以进行药物处理。     

痛风性关节炎动物模型的改良

目的通过改良痛风性关节炎动物模型,制备出更符合人类痛风性关节炎机制的动物模型。方法通过Coderre法与次黄嘌呤相结合（模型B）、Coderre法与氧嗪酸相结合（模型A）的方法建立痛风性关节炎动物模型,采用全自动生化分析仪测定血尿酸水平,观察关节滑膜组织形态学改变及大鼠不同时相步态变化,并将两种方法加以比较。结果模型B组大鼠血尿酸水平显著降低,关节滑膜组织形态学及不同时相大鼠步态改变明显,与模型A组比较有统计学意义。结论Coderre法与次黄嘌呤相结合方法建立大鼠痛风性关节炎动物模型更符合人类痛风性关节炎机制的动物模型。     

去卵巢骨质疏松症大鼠模型骨密度的变化

观察3月龄SD雌鼠切除双侧卵巢29周内模型组和雌二醇组动物第三腰椎及股骨骨密度的变化,同时与假手术组比较。结果表明,SD雌性大鼠切除卵巢9周股骨骨密度明显低于假手术组(P<001),13周后腰椎骨骨密度也有显著差异(P<001),29周内模型稳定。皮下注射苯甲酸雌二醇40μgkg×2次周,在12周能够明显提高切卵巢SD大鼠的股骨骨密度并使其接近正常,16周后第三腰椎的骨密度也能够明显提高。切卵巢骨质疏松症大鼠模型治疗性给药需要大约9～13周左右模型才能成功,切除卵巢后29周内模型仍然稳定。     

Extracorporeal shockwave therapy in osteoporotic osteoarthritis of the knee in rats: an experiment in animals

Introduction

This study investigated the effectiveness of extracorporeal shockwave therapy (ESWT) in osteoporotic (OP) osteoarthritis (OA) of rat knee.

Methods

Fifty-six rats were divided into seven groups including sham, OA, OP, OA + OP, OA + ESWT, OP + ESWT, and OA + OP + ESWT groups. The evaluations included gross pathology, bone mineral density (BMD), micro-computed tomography (micro-CT) scan, bone-strength test, histopathologic examination, and immunohistochemical analysis.

Results

On gross pathology, group OA + OP showed larger areas of osteoarthritic changes than did groups OA and OP, as compared with the sham group. BMD and bone strength significantly decreased in groups OA, OP, and OA + OP relative to the sham group, and ESWT significantly improved BMD and bone-strength changes. On micro-CT scan, the subchondral plate thickness significantly decreased, and the bone porosity increased in groups OA, OP, and OA + OP, and ESWT significantly improved the changes in subchondral-plate thickness and bone porosity. In histopathologic examination, Mankin score and safranin O score significantly increased in groups OA and group OA + OP, but not in group OP relative to the sham group, and ESWT significantly improved the changes. In immunohistochemical analysis, Dickkopf-1 (DKK-1) significantly increased, but vessel endothelial growth factor (VEGF), proliferating cell nuclear antigen (PCNA), and bone morphogenetic protein 2 (BMP-2) decreased in groups OA, OP, and OA + OP relative to the sham group, and ESWT significantly reversed the changes.

Conclusions

Osteoporosis increased the severity of cartilage damage in osteoarthritis of the knee. ESWT showed effectiveness in the reduction of osteoporotic osteoarthritis of the knee in rats.     

去卵巢致骨质疏松症大鼠腺垂体FS细胞中Cx43、S-100蛋白表达--激光扫描共聚焦显微镜观察

检测间隙连接蛋白Cx43、神经组织蛋白S-100在去卵巢致骨质疏松症(OVX-OP)大鼠腺垂体滤泡星形细胞(FS细胞)中的表达.实验采用10月龄未孕产SD雌性大鼠40只,随机均分为卵巢切除组(OVX组,n=20)和假性手术对照组(Sham组,n=20).于术后6周末用双能X线骨吸收测量法(DEXA)测量大鼠全身及腰椎4-6(L4-6)骨密度(BMD).取两组大鼠垂体,制成连续切片.应用FITC标记的IgG探针,对腺垂体组织中Cx43和S-100进行间接免疫荧光染色,并用激光扫描共聚焦显微镜(LSCM)定位和定量分析腺垂体FS细胞中Cx43、S-100的表达.结果发现,术后6周末OVX组大鼠全身及L4-6BMD均明显低于Sham组值(P<0.01,P<0.01).Cx43阳性荧光反应主要定位于相邻的FS细胞的胞浆中和/或胞膜上.OVX组Cx43阳性表达荧光强度和表达阳性率均显著低于Sham组(P<0.01).S-100蛋白表达定位于FS细胞的胞浆中,两组间S-100阳性表达荧光强度和表达阳性率无显著差异(P>0.05).本研究提示,SD大鼠OVX术后6周出现骨质疏松变化;OVX大鼠腺垂体FS细胞数量无明显变化、而Cx43表达显著下降,后者的变化可能与大鼠OVX-OP发生相关.     

Pulsed electromagnetic fields prevent osteoporosis in an ovariectomized female rat model: a prostaglandin E2-associated process

With the use of Helmholtz coils and pulsed electromagnetic field (PEMF) stimulators to generate uniform time varying electromagnetic fields, the effects of extremely low frequency electromagnetic fields on osteoporosis and serum prostaglandin E(2) (PGE(2)) concentration were investigated in bilaterally ovariectomized rats. Thirty-five 3 month old female Sprague-Dawley rats were randomly divided into five different groups: intact (INT), ovariectomy (OVX), aspirin treated (ASP), PEMF stimulation (PEMF + OVX), and PEMF stimulation with aspirin (PEMF + ASP) groups. All rats were subjected to bilateral ovariectomy except those in INT group. Histomorphometric analyses showed that PEMF stimulation augmented and restored proximal tibial metaphyseal trabecular bone mass (increased hard tissue percentage, bone volume percentage, and trabecular number) and architecture (increased trabecular perimeter, trabecular thickness, and decreased trabecular separation) in both PEMF + OVX and PEMF + ASP. Trabecular bone mass of PEMF + OVX rats after PEMF stimulation for 30 days was restored to levels of age matched INT rats. PEMF exposure also attenuated the higher serum PGE(2) concentrations of OVX rats and restored it to levels of INT rats. These experiments demonstrated that extremely low intensity, low frequency, single pulse electromagnetic fields significantly suppressed the trabecular bone loss and restored the trabecular bone structure in bilateral ovariectomized rats. We, therefore, conclude that PEMF may be useful in the prevention of osteoporosis resulting from ovariectomy and that PGE(2) might relate to these preventive effects.     

Periodontal Regeneration Using Strontium-Loaded Mesoporous Bioactive Glass Scaffolds in Osteoporotic Rats

Recent studies demonstrate that the rate of periodontal breakdown significantly increased in patients compromised from both periodontal disease and osteoporosis. One pharmacological agent used for their treatment is strontium renalate due to its simultaneous ability to increase bone formation and halt bone resorption. The aim of the present study was to achieve periodontal regeneration of strontium-incorporated mesoporous bioactive glass (Sr-MBG) scaffolds in an osteoporotic animal model carried out by bilateral ovariectomy (OVX). 15 female Wistar rats were randomly assigned to three groups: control unfilled periodontal defects, 2) MBG alone and 3) Sr-MBG scaffolds. 10 weeks after OVX, bilateral fenestration defects were created at the buccal aspect of the first mandibular molar and assessed by micro-CT and histomorphometric analysis after 28 days. Periodontal fenestration defects treated with Sr-MBG scaffolds showed greater new bone formation (46.67%) when compared to MBG scaffolds (39.33%) and control unfilled samples (17.50%). The number of TRAP-positive osteoclasts was also significantly reduced in defects receiving Sr-MBG scaffolds. The results from the present study suggest that Sr-MBG scaffolds may provide greater periondontal regeneration. Clinical studies are required to fully characterize the possible beneficial effect of Sr-releasing scaffolds for patients suffering from a combination of both periodontal disease and osteoporosis.     

Effect of low‐level laser therapy and oxytocin on osteoporotic bone marrow‐derived mesenchymal stem cells

Postmenopausal osteoporosis (OP) is a major concern for public health. Low‐level laser therapy (LLLT) has a positive effect on the health of bone marrow mesenchymal stem cells (BMMSCs). The purpose of this study is to evaluate the influence of LLLT and oxytocin (OT) incubation—individually and in combination—on osteoporotic BMMSCs in ovariectomized rats. Twelve female rats were randomized into two groups to undergo either a sham surgery (sham group) or ovariectomy‐induced osteoporosis (OVX group). MSCs harvested from the BM of healthy and OVX rats underwent culture expansion. There were five groups. In Groups one (sham‐BMMSC) and two (OVX‐BMMSC) the cells were held in osteogenic condition medium without any intervention. In the group three (OT), OT incubation with optimum dose was performed for 48 h (two times, 10^?12 molar). In Group four, laser‐treated‐OVX‐BMMSCs were treated with optimum protocol of LLLT (one time, 1.2 J/cm²). In Group five (laser + OT group), the OT incubation plus the laser irradiation was performed. The biostimulatory effect of LLLT is demonstrated by a significant increase in the viability of OVX‐BMMSCs, cell cycle, and extracellular levels of Transforming growth factor beta (TGF‐β), insulin‐like growth factor‐I (IGF‐I), and Alkaline phosphatase (ALP) compared to control OVX‐BMMSCs and/or the sham group. OT incubation and laser + OT incubation have a positive effect on OVX‐BMMSCs. However, LLLT is more effective statistically. We conclude that LLLT significantly improved cell viability, enhanced the osteogenic potential of the OVX‐BMMSCs, and increased the extracellular levels of the TGF‐β, IGF‐I, and ALP.     

Subchondral bone microstructural damage by increased remodelling aggravates experimental osteoarthritis preceded by osteoporosis

Introduction  

Osteoporosis (OP) increases cartilage damage in a combined rabbit model of OP and osteoarthritis (OA). Accordingly, we assessed whether microstructure impairment at subchondral bone aggravates cartilage damage in this experimental model.     

骨质疏松兔松质骨微观结构和微观成分的时间序贯性变化

目的:研究去势手术建立骨质疏松兔模型中松质骨微观结构和微观成分的时间序贯性变化。方法:40只新西兰白兔随机分为假手术组(sham组,n=20)和骨质疏松组(OP组,n=20)。OP组兔子给予去势手术处理,sham组给予假手术处理。分别于术后的0周、4周、6周、8周,利用DXA测量腰椎骨密度(每组每个时间点选择5只动物)。之后处死动物,采集腰椎标本。利用Micro-CT、FTIR、腰椎轴向压缩试验得到松质骨的微观结构、微观成分(骨矿盐晶体和胶原)和宏观力学参数。利用t检验比较同一时间点两组之间的相关参数。结果:OP组BMD逐渐下降,松质骨微观结构逐渐疏松,微观组成属性逐渐改变,宏观力学强度均逐渐下降。FTIR在4周时即检测到OP组腰椎骨矿盐和胶原基质比(P=0.046)、骨矿盐结晶度(P=0.018)、胶原交联比(P=0.006)发生显著性改变,早于BMD和微观结构的变化。OP组腰椎宏观生物力学强度在第8周时达到最低点(P=0.001)。结论:去势手术后,腰椎松质骨骨矿盐晶体和胶原属性最早发生变化,松质骨微观成分和微观结构的改变是导致椎体强度明显改变的原因。FTIR技术可以较早的检测到骨质疏松发生过程中骨组织微观成分的改变。     

Effects of Multi-Deficiencies-Diet on Bone Parameters of Peripheral Bone in Ovariectomized Mature Rat

Many postmenopausal women have vitamin D and calcium deficiency. Therefore, vitamin D and calcium supplementation is recommended for all patients with osteopenia and osteoporosis. We used an experimental rat model to test the hypothesis that induction of osteoporosis is more efficiently achieved in peripheral bone through combining ovariectomy with a unique multi-deficiencies diet (vitamin D depletion and deficient calcium, vitamin K and phosphorus). 14-week-old Sprague-Dawley rats served as controls to examine the initial bone status. 11 rats were bilaterally ovariectomized (OVX) and fed with multi-deficiencies diet. Three months later the treated group and the Sham group (n = 8) were euthanized. Bone biomechanical competence of the diaphyseal bone was examined on both, tibia and femur. Image analysis was performed on tibia via µCT, and on femur via histological analysis. Lower torsional stiffness indicated inferior mechanical competence of the tibia in 3 month OVX+Diet. Proximal metaphyseal region of the tibia showed a diminished bone tissue portion to total tissue in the µCT despite the increased total area as evaluated in both µCT and histology. Cortical bone showed higher porosity and smaller cross sectional thickness of the tibial diaphysis in the OVX+Diet rats. A lower ALP positive area and elevated serum level of RANKL exhibited the unbalanced cellular interaction in bone remodeling in the OVX+Diet rat after 3 month of treatment. Interestingly, more adipose tissue area in bone marrow indicated an effect of bone loss similar to that observed in osteoporotic patients. Nonetheless, the presence of osteoid and elevated serum level of PTH, BGP and Opn suggest the development of osteomalacia rather than an osteoporosis. As the treatment and fracture management of both osteoporotic and osteomalacia patients are clinically overlapping, this study provides a preclinical animal model to be utilized in local supplementation of minerals, drugs and growth factors in future fracture healing studies.